| Every year, about 200 thousand women die from uterine cervical cancer worldwidely. The uterine cervical cancer is particularly common in developing countries, and is one of the three gynecological malignant tumors. Uterine cervical squamous cell carcinoma accounts for about 90% of uterine cervical cancer, and its incidence and mortality still maintain a high level. However, its mechanism is still not clear. The tumorigeness of uterine cervical squamous cell carcinoma undergoes a series of complicated changes, developing the malignant phenotype ultimately. In the development of malignant transformation, there are many changes in gene and protein, disorders of cervical epithelial cell differentiation, abnormal hyperplasia, cytokines regulating gowth and development, and so on, which lead to tumor formation finaly.Annexin superfamily contains A, B, C, D, E five families, in which family A was expressed in vertebrate cells. They are similar not only in structure, but also in function, for example, they are involved in activities of the cytoskeleton structure, cell pinocytosis and exocytosis, cell membrane receptor conformation and function of regulation, regulation of ion channels and other important functions. Many foreign scholars found that the expression of family A members was different between certain tumor tissues and normal tissues. Human AnnexinA7 gene is located in chromosome 10q21 with 14 exons, encoding two subtypes, and their molecular weights are 47 kD and 51 kD. In other tissues, only the 47 kDa isoform is expressed, except in heart and brain where both isoforms are present. Up to now, there is no conclusive research findings on the functions of AnnexinA7. The reports are not consistent in experiments, and most of its research are simply stuck in the stage of speculation and hypothesis. In this study, human uterine cervical squamous cell carcinoma tissues and normal tissues were collected to detect the expression of AnnexinA7. The results showed that the expression of AnnexinA7 was up-regulated in human uterine cervical squamous carcinoma tissue and it may play an important role in tumorigeness of uterine cervical squamous cell carcinoma. In order to further study the mechanism of AnnexinA7, we first cultured human cervical carcinoma cell line HeLa. When the effection of the siRNA was assured, the siRNA was transfected into Hela cells to knockdown the expression of AnnexinA7. After finding that the knockdown of AnnexinA7 promotes theapoptosis of Hela cells, siRNA was used again to knockdown the expression of Annexin A7. At last, immunohistochemistry and Western blot were used to detect the expression of Bax, Bcl-2, Fas, Fasl and Apaf-1 in Hela cells. In our study, we preliminarily study the effection of AnnexinA7 on the apoptosis of Hela cells and the possible apoptotic pathway to establish the foundation for further studying on the mechanism of tumorigeness.Partâ… The expression of Annexin A7 in human uterine cervical Squamous carcinoma and normal tissuesObjective: To detect the expression of Annexin A7 in human uterine cervical squamous cell carcinoma tissues and normal tissues.Methods:1 Specimen collection: The fresh cervical squamous cell carcinoma and normal cervical tissues were collected from Hebei Medical University Fourth Hospital and The People's Hospital of Hebei Province, each 25 cases stored in liquid nitrogen. The paraffin imbedded specimen of cervical squamous cell carcinoma and normal cervical tissues were obtained from The People's Hospital of Hebei Province, each 25 cases placed in 4oC refrigerator.2 Immunohistochemistry was used to detect the differential expression of AnnexinA7 in human uterine cervical squamous carcinoma and normal tissues.3 Western blotting was used to observe the differential expression of Annexin A7 between human uterine cervical squamous cell carcinoma and normal tissues.4 RT-PCR was used to observe the differential expression of AnnexinA7 in human uterine cervical squamous carcinoma and normal tissues.Results:1 The IHC result showed that the expression of Annexin A7 was significantly increased in the tumor tissues(P<0.05).2 The expression of annexin A7 was significantly increased in the tumor tissues at the protein level(P<0.05).3 The expression of annexin A7 was significantly increased in the tumor tissues at the mRNA level(P<0.05).Conclusion:1 The expression of annexin A7 is up-regulated in human uterine cervical squamous carcinoma tissue.2 Annexin A7 was involved in tumorigeness of uterine cervical squamous cell carcinoma.Partâ…¡Study on the influence of Annexin A7 knockdown on proliferation and apoptosis of HeLa CellsObjective: To study whether the knockdown of Annexin A7 can affect apoptosis and proliferation of HeLa cells.Methods:1 Immunohistochemistry and Western blot were used to identify the expression of annexin A7 in human cervical carcinoma cell line HeLa.2 Western blot was used to identify the inhibitory effects of siRNA interference on the expression of annexin A7.3 MTT assay was conducted to detect the impact on the proliferation of HeLa cells after the expression of annexin A7 was inhibited by siRNA.4 Flow cytometry and Confocal laser were conducted to detect the impact on the apoptosis of HeLa cells after the expression of annexin A7 was inhibited by siRNA.Results:1 Annexin A7 was expressed in Hela cells. 2 The ANXA7 expression of siRNA group was decreased significantly (P<0.05), compared with the negative control group and blank control group. The ANXA7 expression of negative control group showed no significant difference compared with the blank control group (P>0.05).3 The cell proliferation of ANXA7siRNA group showed no significant difference compared with the negative control group and blank control group (P>0.05).4 The cells apoptotic rate of NXA7siRNA group was significantly increased(P<0.05) compared with the negative control group and blank control group. The cells apoptotic rate of negative control group and blank control group showed no significant difference (P>0.05).5 The cell apoptosis of ANXA7siRNA group was increased significantly(P<0.05), compared with the negative control group and blank control group. The cell apoptosis of negative control group and blank control group showed no significant difference(P>0.05).Conclusion:1 The knockdown of Annexin A7 may not affect the proliferation of HeLa cells.2 The knockdown of Annexin A7 promotes the apoptosis of Hela cells.3 Annexin A7 may inhibit apoptosis of Hela cells.Partâ…¢Study on the preliminary mechanism of the effects of Annexin A7 on apoptosis of Hela cellsObjective: To detect the possible mechanism of the effects of Annexin A7 on apoptosis of Hela cells.Methods:1 Immunohistochemistry was used to detect the expression of Bax, Bcl-2, Fas, Fasl and Apaf-1 in ANXA7siRNA group, negative control group and blank control group.2 Western blot was used to detect the expression of Bax, Bcl-2, Fas, Fasl and Apaf-1 in ANXA7siRNA group, negative control group and blank control group. Results:1 Western blot:(1) The Bax and Fas expression of ANXA7siRNA group was increased significantly(P<0.05), compared with the negative control group and blank control group.(2) The expression of Bcl-2, Fasl and Apaf-1 showed no significant difference (P>0.05) in the three groups(ANXA7siRNA group, negative control group and blank control group).2 Immunohistochemistry:(1) The Bax and Fas expression of ANXA7siRNA group was increased significantly(P<0.05), compared with the negative control group and blank control group.(2) The expression of Bcl-2, Fasl and Apaf-1 showed no significant difference (P>0.05) in the three groups(ANXA7siRNA group, negative control group and blank control group).Conclusion:1 When the expression of AnnexinA7 was suppressed, Bax expression increased in ANXA7siRNA group, indicating that Annexin A7 may be apoptosis gene.2 When the expression of Annexin A7 was suppressed, Fas expression increased in ANXA7siRNA group, indicating that ANXA7 may be through cell death receptor pathway to inhibite the apoptosis of Hela cells.
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